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Human TIMP-2(Tissue Inhibitors of Metalloproteinase 2) ELISA Kit

Citations(6)Uniprot : P16035

Cat.No.:E-EL-H1453
Reactivity: Human

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Product Details

Properties

Assay type	Sandwich-ELISA
Format	96T/48T
Assay time	3.5h
Detection range	0.16-10 ng/mL
Sensitivity	0.10 ng/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 100μL
Specificity	This kit recognizes Human TIMP-2 in samples. No significant cross-reactivity or interference between Human TIMP-2 and analogues was observed.
Reproducibility	Both intra-CV and inter-CV are < 10%.
Application	This ELISA kit applies to the in vitro quantitative determination of Human TIMP-2 concentrations in serum, plasma and other biological fluids.

Test Principle

This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human TIMP-2. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human TIMP-2 and Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human TIMP-2, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Human TIMP-2. You can calculate the concentration of Human TIMP-2 in the samples by comparing the OD of the samples to the standard curve.

Kit components & Storage

An unopened kit can be stored at 2-8℃ for 1 month. If the kit is not supposed to be used within 1 month, store the components separately according to the following conditions once the kit is received.

Item	Specifications	Storage
Micro ELISA Plate(Dismountable)	96T: 8 wells ×12 strips 48T: 8 wells ×6 strips	-20℃, 6 months
Reference Standard	96T: 2 vials 48T: 1 vial
Concentrated Biotinylated Detection Ab (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL
Concentrated HRP Conjugate (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL	-20℃(Protect from light), 6 months
Reference Standard & Sample Diluent	1 vial, 20 mL	2-8°C, 6 months
Biotinylated Detection Ab Diluent	1 vial, 14 mL
HRP Conjugate Diluent	1 vial, 14 mL
Concentrated Wash Buffer (25×)	1 vial, 30 mL
Substrate Reagent	1 vial, 10 mL	2-8℃(Protect from light)
Stop Solution	1 vial, 10 mL	2-8°C
Plate Sealer	5 pieces
Manual	1 copy
Certificate of Analysis	1 copy

Technical Data

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human TIMP-2 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human TIMP-2 were tested on 3 different plates, 20 replicates in each plate.

	Intra-assay Precision			Inter-assay Precision
Sample	1	2	3	1	2	3
n	20	20	20	20	20	20
Mean (ng/mL)	0.50	1.30	4.90	0.50	1.40	5.10
Standard deviation	0.03	0.07	0.24	0.03	0.08	0.17
CV (%)	6.00	5.38	4.90	6.00	5.71	3.33

Recovery

The recovery of Human TIMP-2 spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Sample Type	Range (%)	Average Recovery (%)
Serum(n=8)	92-107	100
EDTA plasma (n=8)	95-109	101
Cell culture media (n=8)	89-101	96

Linearity

Samples were spiked with high concentrations of Human TIMP-2 and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Target Information

Database Links	SwissProt: P16035
Synonyms	TIMP2, CSC-21K
Research Area	Cancer, Cell Biology, Cardiovascular, Signal transduction

Assay Procedures

	1. Add 100μL standard or sample to the wells. Incubate for 90 min at 37°C
	2. Discard the liquid, immediately add 100μL Biotinylated Detection Ab working solution to each well. Incubate for 60 min at 37°C
	3. Aspirate and wash the plate for 3 times
	4. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times
	5. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C
	6. Add 50μL Stop Solution
	7. Read the plate at 450nm immediately. Calculation of the results

Citations

BIOMEDICINE & PHARMACOTHERAPY (2023) IF: 7.419
Stiff substrates inhibit collagen accumulation via integrin α2β1, FAK and Src kinases in human atrial fibroblast and myofibroblast cultures derived from patients with aortal stenosis

DOI: 10.1016/j.biopha.2023.114289

Sample: cardiac fibroblast
Cells (2021) IF: 6.6
The Stiffness of Cardiac Fibroblast Substrates Exerts a Regulatory Influence on Collagen Metabolism via α2β1 Integrin, FAK and Src Kinases

DOI: 10.3390/cells10123506

Sample: cardiac fibroblast
EBioMedicine (2019) IF: 6.68
Three kinds of corneal host cells contribute differently to corneal neovascularization

DOI: 10.1016/j.ebiom.2019.05.026

PMID: 31126890
Sample: Tissue homogenate
Aging-US (2022) IF: 5.682
TIMP-2 regulates 5-Fu resistance via the ERK/MAPK signaling pathway in colorectal cancer

DOI: 10.18632/aging.203793

PMID: 35022331
Sample: serum
RENAL FAILURE (2022) IF: 3.222
Urinary NGAL, IGFBP-7, and TIMP-2: novel biomarkers to predict contrast medium-induced acute kidney injury in children

DOI: 10.1080/0886022X.2022.2075277

PMID: 36120960
Sample: Serial urinary samples
INTERNATIONAL JOURNAL OF EXPERIMENTAL PATHOLOGY (2019) IF: 1.494
MT1‐MMP evaluation in neointimal hyperplasia in the late follow‐up after prosthesis implantation

DOI: 10.1111/iep.12310

Sample: neointima

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